Department of Molecular and Cell Biology, University of California, Berkeley, United States.
Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States.
Elife. 2019 Jan 4;8:e40805. doi: 10.7554/eLife.40805.
Optical imaging has become a powerful tool for studying brains in vivo. The opacity of adult brains makes microendoscopy, with an optical probe such as a gradient index (GRIN) lens embedded into brain tissue to provide optical relay, the method of choice for imaging neurons and neural activity in deeply buried brain structures. Incorporating a Bessel focus scanning module into two-photon fluorescence microendoscopy, we extended the excitation focus axially and improved its lateral resolution. Scanning the Bessel focus in 2D, we imaged volumes of neurons at high-throughput while resolving fine structures such as synaptic terminals. We applied this approach to the volumetric anatomical imaging of dendritic spines and axonal boutons in the mouse hippocampus, and functional imaging of GABAergic neurons in the mouse lateral hypothalamus in vivo.
光学成像是研究活体大脑的有力工具。成年大脑的不透明性使得微内窥镜成为首选方法，其中包括将光学探头（如梯度折射率（GRIN）透镜）嵌入脑组织中以提供光学中继，从而可以对深埋在大脑结构中的神经元和神经活动进行成像。我们将贝塞尔聚焦扫描模块集成到双光子荧光微内窥镜中，从而轴向扩展了激发焦点，并提高了其横向分辨率。通过在 2D 中扫描贝塞尔焦点，我们在实现高分辨率的同时，以高通量的方式对神经元体积进行成像，解析出突触末梢等精细结构。我们将这种方法应用于小鼠海马体树突棘和轴突末梢的容积解剖成像，以及小鼠外侧下丘脑 GABA 能神经元的功能成像。
